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Power-to-X (P2X) technologies will play a more important role in the conversion of electric power to storable energy carriers, commodity chemicals and even food and feed. Among the different P2X technologies, microbial components form cornerstones of individual process steps. This review comprehensively presents the state-of-the-art of different P2X technologies from a microbiological standpoint. We are focusing on microbial conversions of hydrogen from water electrolysis to methane, other chemicals and proteins. We present the microbial toolbox needed to gain access to these products of interest, assess its current status and research needs, and discuss potential future developments that are needed to turn todays P2X concepts into tomorrow's technologies.

Fermentation at elevated hydrostatic pressure is a novel strategy targeting product selectivity. However, the role of inoculum history and cross-resistance, that is, acquired tolerance from incubation under distinctive environmental stress, remains unclear in high-pressure operation. In our here presented work, we studied fermentation and microbial community responses of halotolerant marine sediment inoculum (MSI) and anaerobic digester inoculum (ADI), pre-incubated in serum bottles at different temperatures and subsequently exposed to mild hydrostatic pressure (MHP

Background Elevated CO2 partial pressure (pCO2) has been proposed as a potential steering parameter for selective carboxylate production in mixed culture fermentation. It is anticipated that intermediate product spectrum and production rates, as well as changes in the microbial community, are (in)directly influenced by elevated pCO2. However, it remains unclear how pCO2 interacts with other operational conditions, namely substrate specificity, substrate-to-biomass (S/X) ratio and the presence of an additional electron donor, and what effect pCO2 has on the exact composition of fermentation products. Here, we investigated possible steering effects of elevated pCO2 combined with (1) mixed substrate (glycerol/glucose) provision; (2) subsequent increments in substrate concentration to increase the S/X ratio; and (3) formate as an additional electron donor. Results Metabolite predominance, e.g., propionate vs. butyrate/acetate, and cell density, depended on interaction effects between pCO2–S/X ratio and pCO2–formate. Individual substrate consumption rates were negatively impacted by the interaction effect between pCO2–S/X ratio and were not re-established after lowering the S/X ratio and adding formate. The product spectrum was influenced by the microbial community composition, which in turn, was modified by substrate type and the interaction effect between pCO2–formate. High propionate and butyrate levels strongly correlated with Negativicutes and Clostridia predominance, respectively. After subsequent pressurized fermentation phases, the interaction effect between pCO2–formate enabled a shift from propionate towards succinate production when mixed substrate was provided. Conclusions Overall, interaction effects between elevated pCO2, substrate specificity, high S/X ratio and availability of reducing equivalents from formate, rather than an isolated pCO2 effect, modified the proportionality of propionate, butyrate and acetate in pressurized mixed substrate fermentations at the expense of reduced consumption rates and increased lag-phases. The interaction effect between elevated pCO2 and formate was beneficial for succinate production and biomass growth with a glycerol/glucose mixture as the substrate. The positive effect may be attributed to the availability of extra reducing equivalents, likely enhanced carbon fixating activity and hindered propionate conversion due to increased concentration of undissociated carboxylic acids.

Anaerobic digestion of a mono-feedstock often causes low methane yields and process instability. An effective strategy to overcome these barriers is co-digestion with animal manure. The obtained process improvement is often attributed to buffer capacity, nutrients, vitamins and trace metals, and microorganisms present in manure, but it remains unknown which factor plays the key role in digester performance. Here, we investigated anaerobic digestion of cocoa waste in four different treatments: mono-digestion, addition of synthetic nutrients, co-digestion with sterile cow manure, and co-digestion with raw cow manure. Co-digestion with raw manure resulted in the highest methane yield of 181 ± 39 L kg−1 VS (volatile solids), similar to the co-digestion with sterile manure, i.e., 162 ± 52 L kg−1 VS. The supplementation of synthetic nutrients to the anaerobic digestion of cocoa waste only temporarily increased methane yield, indicating that this will tackle a lack of nutrients in the short term, but has a limited long-term contribution to the stabilization of the process. Hence, because of the inability of synthetic nutrients to stabilize the digestion process and the similarity between the digesters fed sterile and raw manure, both at the physico-chemical and microbial level, the key contribution of manure co-digestion with cocoa seems to be the provision of buffering capacity.

Microorganisms in anaerobic digestion (AD) are essential for wastes/pollutants treatment and energy recovery. Due to microbial enormous diversity, developing effective perspectives to understand microbial roles therein is urgent. This study conducted AD of swine manure, used an ensemble-based network analysis to distinguish interconnected, unconnected, copresence (positively interconnected) and mutual-exclusion (negatively inter-connected) microorganisms within microbial communities, and explored their importance towards AD perfor-mances, using amplicon sequencing of 16S rRNA and 16S rRNA gene. Our analyses revealed greater importance of interconnected than unconnected microorganisms towards CH4 production and AD multifunctionality, which was attributed to higher niche breadth, deterministic community assembly, community stability and phyloge-netic conservatism. The diversity was higher in unconnected than interconnected microorganisms, but was not linked to AD performances. Compared to copresence microorganisms, mutual-exclusion microorganisms showed greater and equal importance towards CH4 production and AD multifunctionality, which was attributed to their roles in stabilizing microbial communities. The increased feedstock biodegradability, by replacing part of manure with fructose or apple waste, hardly affected the relative importance of interconnected versus unconnected microorganisms towards CH4 production or AD multifunctionality. Our findings develop a new framework to understand microbial roles, and have important implications in targeted manipulation of critical microorganisms in waste-treatment systems.

Summary Pure (single) cultures of microorganisms and mixed microbial communities (microbiomes) have been important for centuries in providing renewable energy, clean water and food products to human society and will continue to play a crucial role to pursue the Sustainable Development Goals. To use microorganisms effectively, microbial engineered processes require adequate control. Microbial communities are shaped by manageable deterministic processes, but also by stochastic processes, which can promote unforeseeable variations and adaptations. Here, we highlight the impact of stochasticity in single culture and microbiome engineering. First, we discuss the concepts and mechanisms of stochasticity in relation to microbial ecology of single cultures and microbiomes. Second, we discuss the consequences of stochasticity in relation to process performance and human health, which are reflected in key disadvantages and important opportunities. Third, we propose a suitable decision tool to deal with stochasticity in which monitoring of stochasticity and setting the boundaries of stochasticity by regulators are central aspects. Stochasticity may give rise to some risks, such as the presence of pathogens in microbiomes. We argue here that by taking the necessary precautions and through clever monitoring and interpretation, these risks can be mitigated., In this review paper, we highlight the impact of stochasticity in single culture and microbiome engineering. Stochasticity may give rise to some risks, such as the presence of pathogens in microbiomes. We argue here that by taking the necessary precautions and through clever monitoring and interpretation, these risks can be mitigated.

Summary Anaerobic digesters produce biogas, a mixture of predominantly CH4 and CO2, which is typically incinerated to recover electrical and/or thermal energy. In a context of circular economy, the CH4 and CO2 could be used as chemical feedstock in combination with ammonium from the digestate. Their combination into protein‐rich bacterial, used as animal feed additive, could contribute to the ever growing global demand for nutritive protein sources and improve the overall nitrogen efficiency of the current agro‐ feed/food chain. In this concept, renewable CH4 and H2 can serve as carbon‐neutral energy sources for the production of protein‐rich cellular biomass, while assimilating and upgrading recovered ammonia from the digestate. This study evaluated the potential of producing sustainable high‐quality protein additives in a decentralized way through coupling anaerobic digestion and microbial protein production using methanotrophic and hydrogenotrophic bacteria in an on‐farm bioreactor. We show that a practical case digester handling liquid piggery manure, of which the energy content is supplemented for 30% with co‐substrates, provides sufficient biogas to allow the subsequent microbial protein as feed production for about 37% of the number of pigs from which the manure was derived. Overall, producing microbial protein on the farm from available methane and ammonia liberated by anaerobic digesters treating manure appears economically and technically feasible within the current range of market prices existing for high‐quality protein. The case of producing biomethane for grid injection and upgrading the CO2 with electrolytic hydrogen to microbial protein by means of hydrogen‐oxidizing bacteria was also examined but found less attractive at the current production prices of renewable hydrogen. Our calculations show that this route is only of commercial interest if the protein value equals the value of high‐value protein additives like fishmeal and if the avoided costs for nutrient removal from the digestate are taken into consideration., Anaerobic digesters are the core engine for a biorefinery producing microbial protein. Either the biogas can be used directly for its production, or via biogas upgrading the separate CO2 can be combined with electrolytic hydrogen to produce more protein.

The integration of conversion processes with anaerobic digestion is key to increase value from agricultural waste, like cocoa pod husks, generated in developing countries. The production of one metric ton of cocoa beans generates some 15 metric tonnes of organic waste that is today underutilized. This waste can be converted into added value products by anaerobic digestion, converting part of the cocoa pods to biogas while releasing nutrients, and pyrolysis. Here, we compared different scenarios for anaerobic digestion/slow pyrolysis integration in terms of product portfolio (i.e., biogas, pyrolysis liquids, biochar and pyrolysis gases), energy balance and potential for chemicals production. Slow pyrolysis was performed at 350 degrees C and 500 degrees C on raw cocoa pod husks, as well as on digestates obtained from mono-digestion of cocoa pod husks and co-digestion with cow manure. Anaerobic digestion resulted in 20 to 25 wt% of biogas for mono and co-digestion, respectively. Direct pyrolysis of cocoa pod husks mainly resulted in biochar with a maximum yield of 48 wt%. Anaerobic digestion induced compositional changes in the resulting biochar, pyrolysis liquids and evolved gases after pyrolysis. Pyrolysis of mono-digestatee.g., resulted in a more energy-dense organic phase, rich in valuable phenolics while poorer in light oxygenates that hold a modest value. Our comparison shows that co-digestion/slow pyrolysis at 500 degrees C and mono-digestion/slow pyrolysis at 350 degrees C both present high-potential biorefinery schemes. They can be self-sustaining in terms of energy, while resulting in high quality biochar for nutrient recycling and/or energy recovery, and/or phenolics-rich pyrolysis liquids for further upgrading into biorefinery intermediates.

Microbial life strategy, reflected by rRNA operon (rrn) copy number, determines microbial ecological roles. However, the relationship between microbial life strategy and the energy and nutrient flux in anaerobic digestion (AD) remains elusive. This study investigated microbial rrn copy number and expression ratio using amplicon sequencing of 16S rRNA gene and 16S rRNA, and monitored CH4 daily production to approximate the status of energy and nutrient flux in semi-continuous AD. A significantly positive correlation between the mean rrn copy number of microbial communities in digestate and CH4 daily production was detected in the control treatment fed swine manure. The reduced feedstock complexity, by replacing parts of swine manure with fructose or apple waste, weakened the correlation. When feedstock complexity was increased again, the correlation was strengthened again. Similar results were detected in mean rrn expression ratio of microbial communities. The responses of mean rrn copy number and expression ratio of communities to feedstock addition differed between the reduced feedstock complexity and the control treatment, as well as between in digestate and in straw. Our findings reveal a novel relationship between microbial community life strategy and the energy and nutrient flux, and the roles of feedstock characteristics therein in AD.

Returning the carbonated biomass ash into the soil can enhance the carbon fixation capacity of crop-soil ecosystem, contributing to the achievement of "4 per mille Soils for Food Security and Climate' goal launched at COP21. The CO2 sequestration of crop- soil ecosystem induced by biomass ash, however, highly depended on the soil pH and crops growth, which have not been elucidated. Here, we explored the effects of different management strategies (watering, chemical fertilizer, and CO2-rich biogas slurry on the CO2 fixation performance of tomato ecosystem (Tom-E) containing one tomato plant, biomass ash, and soil, and evaluated the impact of tomato cultivation concerning carbon fixation. Results showed that when burying biomass ash into the soil, biomass ash did not cause physiological stress on tomato growth, and even improved the tomato qualities. Furthermore, through sequestrating CO2 from biomass ash, CO2 fixation capacity of the tomato ecosystem reached about 8.8 +/- 1.10, 13.7 +/- 0.74, and 20.7 +/- 0.53 mmol-CO2/Tom-E for watering, chemical fertilizer, and CO2-rich biogas slurry management strategies, respectively. However, due to the influence of fertilization management strategy, about 57.4% +/- 0.09 and 27.3% +/- 0.09 of CO2 sequestrated in the biomass ash returned to the atmosphere (i.e, CO2 loss) for only watering treatment and conventional fertilization (chemical fertilizer) treatment, respectively. Comparatively, as a pH buffering fertilizer, CO2-rich biogas slurry application improved the bicarbonate assimilation environment of tomato, which contributed to reducing the CO2 loss to only 12.5% +/- 0.01. Clearly, adopting the carbonated biomass ash and CO2-rich biogas slurry as fertilizer provides a suitable and potential strategy for the enhancement of soil carbon sink. (C) 2022 The Author(s). Published by Elsevier B.V.

We investigated the gut microbiota of rabbit fish larvae at three locations in Vietnam (ThuanAn-northern, QuangNam-intermediate, BinhDinh-southern sampling site) over a three-year period. In the wild, the first food for rabbit fish larvae remains unknown, while the juveniles and adults are herbivores, forming schools near the coasts, lagoons, and river mouths, and feeding mainly on filamentous algae. This is the first study on the gut microbiota of the wild fish larvae and with a large number of individuals analyzed spatially and temporally. The Clostridiales order was the most predominant in the gut, and location-by-location alpha diversity showed significant differences in Chao-1, Hill number 1, and evenness. Analysis of beta diversity indicated that the location, not year, had an effect on the composition of the microbiota. In 2014, the gut microbiota of fish from QuangNam was different from that in BinhDinh; in 2015, the gut microbiota was different for all locations; and, in 2016, the gut microbiota in ThuanAn was different from that in the other locations. There was a time-dependent trend in the north-south axis for the gut microbiota, which is considered to be tentative awaiting larger datasets. We found limited variation in the gut microbiota geographically and in time and strong indications for a core microbiome. Five and fifteen OTUs were found in 100 and 99% of the individuals, respectively. This suggests that at this life stage the gut microbiota is under strong selection due to a combination of fish-microbe and microbe-microbe interactions. ispartof: MICROBIAL ECOLOGY vol:79 issue:3 pages:539-551 ispartof: location:United States status: published

Turning raw biogas into biomethane as energy carrier requires the selective removal of CO2 in a biogas upgrading process or a total conversion of CO2 to CH4 which is generally energy intensive. During membrane electrolysis, electrical energy can be used to simultaneously remove CO2 (and H2S) and produce H2 as side product. Biogas is thus scrubbed with catholyte and the captured HCO3− and HS− migrate towards the anode. Simultaneously, cathodic H2 mixes with residual biogas in a ratio that can be fine-tuned. We obtained in one step an ideal 4:1 H2:CO2 ratio in the reactor off gas. Subsequently the gas could be further upgraded via chemoautotrophic microbial conversion of CO2 to CH4. Biomethanation delivered biomethane with 98.9 ± 0.9% purity. The electrochemically-assisted scrubbing and stripping of CO2 and H2S resulted in high CO2 removal efficiencies (up to 100%), without addition of chemicals. The system was flexible depending on temporarily available power. Electrochemical biogas upgrading (EBU) can be envisaged as a scalable and decentralized storage of excess or off-peak renewable power, making better use of the power input used to drive a biological CO2 conversion.

Source-separated urine is of interest for nutrient recovery. Most nitrogen recovery technologies rely on ammonia (NH3) as input, which requires ureolysis. As urease positive bacteria are widespread, source-separated urine is unstable, not only leading to NH3 release but also loss, odor nuisance, and downstream scaling. Hence, ureolysis ideally occurs in a closed controlled environment close to the toilet. We characterized microbial-induced ureolysis, subsequent divalent cation precipitation, and fermentation in anaerobic sequencing batch reactors (SBRs) at 15 °C and 28 °C. Temperatures were a proxy for urine hydrolysis in a wet well at street level or in the toilet, respectively. The need for inoculation and the metabolic stability was assessed by inoculation with autofermented urine or a mixture of anaerobic digestion and fermentation sludge. The highest specific ureolysis rates in the SBRs were achieved at 28 °C: 2107 ± 395 and 1948 ± 1121 mg N g VSS-1 d-1, for the mixed and autofermented inoculum, respectively. For Ca2+ and Mg2+ precipitation, and organics fermentation, autofermented urine at 28 °C performed best with 47.9 ± 16.4 mg Ca2+ g VSS-1 d-1, 8.2 ± 4.6 mg Mg2+ g VSS-1 d-1, and 623 ± 129 mg VFA-COD g VSS-1 d-1, respectively. This indicates the hydrolysis reactor should be close to the toilet. The selected inoculum did not impact ureolysis, whereas both Ca2+ and Mg2+ precipitation and fermentation were better in the SBRs with autofermented urine. Ureolysis was identified as the only process significantly impacting the microbial community, indicating external inoculation would not be required. A urine hydrolysis reactor in the toilet without external inoculation could thus serve as a controlled environment to release NH3 and remove divalent cations to prevent scaling in downstream transport and processing. For practical implementation in a household toilet, the reactor should be designed for user-friendly precipitate discharge and odor control. ispartof: WATER RESEARCH vol:148 pages:97-105 ispartof: location:England status: published

To adhere to the Paris Agreement of 2015, we need to store several Gigatonnes (Gt) of carbon annually. In the last years, a variety of technologies for carbon capture and storage (CCS) and carbon capture and usage (CCU) have been demonstrated. While conventional CCS and CCU are techno-economically feasible, their climate change mitigation potentials are limited, due to limited amount of CO2 that can be captured. Hence, there is an urgent need to explore other CCS and CCU routes. Here we discuss an interesting alternative route for capture of carbon dioxide from industrial point sources, using CO2-binding, so-called autotrophic aerobic bacteria to produce microbial biomass as a C-storage product. The produced microbial biomass is often referred to as microbial protein (MP) because it has a crude protein content of ~70-75%. Depending on the industrial production process and final quality of the produced MP, it can be used for human consumption as meat replacement, protein supplement in animal diets, or slow-release organic fertilizer thus providing both organic nitrogen and carbon to agricultural soils. Here, we discuss the potentials and limitations of this so far unexplored CCU approach. A preliminary assessment of the economic feasibility of the different routes for CO2 carbon avoidance, capture and utilization indicates that the value chain to food is becoming attractive and that the other end-points warrant close monitoring over the coming years. ispartof: SCIENCE OF THE TOTAL ENVIRONMENT vol:644 pages:1525-1530 ispartof: location:Netherlands status: published

The production of shrimp is often performed in earthen outdoor ponds in which the input of feed and faeces on the bottom results in deterioration of the water quality that results in a negative impact on the aquatic animals and the environment. Nitrate application in shrimp ponds could improve pond water and sediment quality by oxidizing anaerobic zones of the sediment. This study was conducted to evaluate the possible benefits of nitrate treatment in combination with probiotics application to decrease sulphide formation in shrimp ponds, which is a crucial problem during the growth phase of shrimp. Three sets of experiments were conducted to describe the mechanism of nitrate treatment: a dual dose nitrate treatment at the start and after 14 days that investigated nitrate treatment with concentrations of 50, 100 and 200 mg/L NO3−, a multiple dose nitrate treatment where lower doses of nitrate (12.5, 25 and 50 mg/L NO3−) were tested with more frequent application (every 4 days) and a microbially assisted nitrate treatment in which 50 mg/L NO3− treatment was applied every 4 days with and without presence of a single probiotic application. A batch system including shrimp feed and faeces, representing the bottom waste produced during 90 days of shrimp culture, was used to test the effect of nitrate addition under anaerobic conditions that mimic the dead zones in shrimp pond bottoms where high amounts of organic matter accumulate. Nitrate addition acted as a curative solution for hydrogen sulphide (H2S) accumulation, rather than preventive. Nitrate amended treatments (12.5 mg/L.day NO3) decreased the accumulated H2S in the headspace of incubation bottles by 93 ± 3% compared to the controls. Nitrate addition increased pH and decreased organic acids accumulation to trace amounts. To justify the decrease in H2S concentration apart from the effect sulphur speciation through pH increase, a separate test was conducted to compare the effect of NaOH with nitrate treatment, in which nitrate treatment resulted in a 62.8% higher decrease in H2S concentrations than the NaOH treatment. The use of a denitrifying strain did not improve the beneficial effect of nitrate, indicating that intrinsic capacity for anoxic conversion of waste by the present community was sufficient. In conclusion, nitrate treatment showed a potential to control H2S accumulation in the shrimp ponds that is a critical problem during the growth of shrimp.

Strategies to enhance process performance of anaerobic digestion remain of key importance to promote wider usage of this technology for integrated resource recovery from organic waste streams. Continuous inoculation of the microbial community in the digester via the feedstock could be such a cost-effective strategy. Here, anaerobic digestion of fresh waste activated sludge (WAS) was compared with sterilized WAS in response to two common process disturbances, i.e. organic overloading and increasing levels of salts, to determine the importance of feedstock inoculation. A pulse in the organic loading rate severely impacted process stability of the digesters fed sterile WAS, with a 92 ± 45% decrease in methane production, compared to a 42 ± 31% increase in the digesters fed fresh WAS, relative to methane production before the pulse. Increasing salt pulses did not show a clear difference in process stability between the digesters fed fresh and sterile WAS, and process recovery was obtained even at the highest salt pulse of 25 g Na+ L-1. Feedstock sterilization through thermal pretreatment strongly impacted the microbial community in the digesters. In conclusion, feedstock thermal pretreatment strongly impacted anaerobic digestion process stability, due to feedstock inoculation and compositional modification. ispartof: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY vol:104 issue:8 pages:3675-3686 ispartof: location:Germany status: published

The mechanisms underlying the advanced performance in anaerobic co-digestion (AcoD) are crucial but remain elusive. This study conducted AcoD of swine manure, rice straw and apple waste (AW, mainly consisting of labile carbon) or fructose (a pure labile carbon), and monitored microbial community abundances, activities and transcriptional profiles in the digestate and on straw. The transformation efficiencies of manure (not straw) to CH4 were promoted in AcoD co-fed manure and AW (by 39 +/- 13%) or fructose (by 65 +/- 14%), compared to the control mono-fed manure, implying labile carbon could trigger a priming effect underlying AcoD advantage. Although digestate-associated and straw-associated communities existed in a same bioreactor, the priming effect mainly linked to the former and was attributed to enhancements in deterministic turnover of active communities, in activities of Firmicutes taxa involved in substrate hydrolysis, and in acetoclastic methanogenesis. These findings provide novel insights to elaborate AcoD processes.

The production of shrimp is often performed in earthen outdoor ponds in which the high input of feed and faeces on the bottom can result in deterioration of the water quality, which negatively impacts the animals and the environment. Here, we investigate the potential of sodium molybdate (Na₂MoO₄·2H₂O), sodium nitrate (NaNO3) and sodium percarbonate (Na2CO3·1.5H2O2) to control sulphide production in a simulated shrimp pond bottom system that included the sediment, overlaying artificial seawater and organic matter input in the form of shrimp feed and shrimp faeces. Sediment depth gradient measurements of oxygen, H2S and pH were obtained during 7 days of incubation using microelectrodes. The most significant impact in terms of H2S, was observed for 50 mg/L sodium molybdate. At the water-sediment interface, there was up to 73% less H2S detected for this treatment in comparison to a control treatment, while in the deeper layers of the sediment it was up to 47% less H2S. The residual sulphate concentrations in the molybdate treated samples were 16 ± 4% higher than the control, indicating an inhibition in sulphate reduction. Nitrate and sodium percarbonate treatments also showed a limited capacity to decrease H2S entering in the water column, yet no clear difference in H2S concentrations in the sediment compared to the control were observed. Molybdate treatment appears to work through the inhibition of sulphate reducing bacteria in situ for the control of H2S production better than the chemical oxygen boosters or nitrate treatment.

The aim of this work was to evaluate the bioenergy potential of cocoa residue via anaerobic digestion. Batch and fed-batch lab-scale reactors were operated under low and high solids conditions. In the batch tests, 59 ± 4% of Chemical Oxygen Demand (COD) was recovered as methane. This corresponded with an average methane yield of 174 (wet) and 193 (dry) L kg-1 volatile solids fed, whereas a series of fed-batch reactors produced 70 ± 24 (wet) and 107 ± 39 (dry) L CH4 kg-1 volatile solids fed during stable conditions. A case study was developed for canton Balao (Ecuador) based on our experimental data, operational estimates and available cocoa waste in the area. Annually, 8341 MWh could be produced, meeting 88% of the current electricity demand in Balao. This case study proves the potential for cocoa waste as a source of renewable energy in rural areas. ispartof: BIORESOURCE TECHNOLOGY vol:265 pages:568-572 ispartof: location:England status: published

Background Early-life antibiotic administration is known to affect gut microbiota and host adiposity, but the effects of antibiotic exposure on skeletal muscle properties remain unknown. The present study evaluated the changes in skeletal muscle properties including myofiber characteristics and composition, as well as intramuscular fat (IMF) content in skeletal muscle of piglets when exposed to a tylosin-containing diet. Results A total of 18 piglets (28 days of age) were randomly allocated into two groups: control basal diet (Control) and Control + 100 mg tylosin phosphate/kg of feed (Antibiotic). The trial lasted for 39 days. High-throughput amplicon sequencing revealed that no significant difference in initial gut microbiota composition was existed between Control and Antibiotic groups. Antibiotic administration increased body weight and growth rate and decreased feed to gain ratio of pigs (P P longissimus muscle (P longissimus muscle was increased by antibiotic exposure (P P Conclusion These results confirm that the growth performance, myofiber composition and muscle lipid metabolism are affected by antibiotic administration, which may be associated with an altered gut microbiota, suggesting that the gut microbiota could be served as a potential target for modulating skeletal muscle properties of host.

High-rate activated sludge (HRAS) systems typically generate diluted sludge which requires further thickening prior to anaerobic digestion (AD), besides the need to add considerable coagulant and flocculant for the solids separation. As an alternative to conventional gravitational settling, a dissolved air flotation (DAF) unit was coupled to a HRAS system or a high-rate contact stabilization (HiCS) system. The HRAS-DAF system allowed up to 78% removal of the influent solids, and the HiCS-DAF 67%. Both were within the range of values typically obtained for HRAS-settler systems, albeit at a lower chemical requirement. The separated sludge had a high concentration of up to 47 g COD L-1, suppressing the need of further thickening before AD. Methanation tests showed a biogas yield of up to 68% on a COD basis. The use of a DAF separation system can thus enable direct organics removal at high sludge concentration and with low chemical needs. ispartof: BIORESOURCE TECHNOLOGY vol:291 ispartof: location:England status: published

The 'microbiome' has become a buzzword. Multiple new technologies allow to gather information about microbial communities as they evolve under stable and variable environmental conditions. The challenge of the next decade will be to develop strategies to compose and manage microbiomes. Here, key aspects are considered that will be of crucial importance for future microbial technological developments. First, the need to deal not only with genotypes but also particularly with phenotypes is addressed. Microbial technologies are often highly dependent on specific core organisms to obtain the desired process outcome. Hence, it is essential to combine omics data with phenotypic information to invoke and control specific phenotypes in the microbiome. Second, the development and application of synthetic microbiomes is evaluated. The central importance of the core species is a no-brainer, but the implementation of proper satellite species is an important route to explore. Overall, for the next decade, microbiome research should no longer almost exclusively focus on its capacity to degrade and dissipate but rather on its remarkable capability to capture disordered components and upgrade them into high-value microbial products. These products can become valuable commodities in the cyclic economy, as reflected in the case of 'reversed sanitation', which is introduced here. ispartof: ENVIRONMENTAL MICROBIOLOGY vol:20 issue:6 pages:1991-2000 ispartof: location:England status: published

Direct anaerobic treatment of domestic wastewater is becoming attractive as it can change a wastewater treatment plant from energy consuming to energy producing. A pilot scale UASB-digester was studied to treat domestic wastewater at temperatures of 10–20 °C and an HRT of 6 h. The results show a stable chemical oxygen demand (COD) removal efficiency of 60 ± 4.6% during the operation at 12.5–20 °C. COD removal efficiency decreased to 51.5 ± 5.5% at 10 °C as a result of insufficient methanogenic capacity caused by low temperature and increased influent COD load (from 2.0 g/(L·d) to 3.0 g/(L·d)). Suspended COD removal was 76.0 ± 9.1% at 10–20 °C. Soluble COD removal fluctuated due to variation of the influent COD concentration, but the effluent COD concentration remained 90 ± 23 mg/L at temperatures between 12.5 and 20 °C. The methane production was 39.7 ± 4.4% of the influent COD, which was 80% of influent biological methane potential. The specific methanogenic activity of the UASB sludge and the digester sludge was 0.26 ± 0.03 and 0.29 ± 0.03 g CH4 COD/(g VSS d), respectively. The methanogenic community analysis revealed an overall dominance of the acetoclastic Methanosaetaceae and the hydrogenotrophic Methanomicrobiales during the operation between 10–20 °C. The results of the UASB-digester treating domestic wastewater at 10–20 °C as reported in this paper provide support for application of anaerobic domestic wastewater treatment in moderate climate zones.

Methanogenic granules from a sewage-fed 298 day old UASB reactor with a high abundance of Nitrospira sp. were evaluated for simultaneous methanogenesis and nitrification. Batch experiments were conducted under anaerobic and oxygen-limited conditions using either 0.04% (v/v) methanol (C) or ammonium chloride (30–100 mg N L−1) (N) or both (N + C). The results showed that C-utilization preceded N-utilization in the N + C sets, indicating that methanogenesis was independent of ammonia removal. Ammonia removal with nitrate generation under oxygen-limited conditions confirmed the nitrification potential of methanogenic granules. However, under anaerobic conditions, the decrease in the ammonium nitrogen concentration did not result in the stoichiometric increase in the nitrite & nitrate concentrations. Furthermore, hydrogenotrophic methanogens disappeared in the N and N + C sets. H2-mediated denitrification could possibly be attributed to the disappearance of both nitrite or nitrate and hydrogenotrophic methanogens. An increase in the population of Methanosaetaceae compensated for methane production via the acetoclastic route. Thus, simultaneous nitrification & H2-mediated denitrification was confirmed as the plausible route for ammonia removal in the methanogenic granules under anaerobic conditions. This finding of in situ ammonia removal in methanogens has opened new avenues for treating C/N wastewaters.

BACKGROUND Microbial community dynamics during the anaerobic digestion of vinasse has been little studied. However, having knowledge about it is essential for early detection of reactor operational difficulties to apply preventive actions. This research studies the microbial community dynamics in the anaerobic digestion of vinasse, linking to experimental observations about product yields and organic matter degradation. RESULTS Methane and sulfide yields decreased with increasing SO42-/COD ratio, while the fraction of organic matter degraded by sulfate reducing bacteria increased from 4.5 ± 0.3% to 27.1 ± 0.6%. The archaeal community showed that acetoclastic Methanosaetaceae were little affected by the increase of the SO42-/COD ratio, in contrast to the Methanomicrobiales and Methanobacteriales population, which decreased during the experiment. The total bacterial diversity was influenced mainly by substrate composition, showing that the increase of the SO42-/COD ratio above 0.10 shifted the bacterial community to a lower richness. CONCLUSION These results provide knowledge on the dynamics of the microbial communities, which can be useful to control de anaerobic digestion of sulfate-rich vinasses, showing that reactor stability equates to the higher ratios between total methanogens and total bacteria gene copy numbers, whereas operational difficulties can be associated to lower bacterial richness and higher community organization.

The anaerobic digestion microbiome has been puzzling us since the dawn of molecular methods for mixed microbial community analysis. Monitoring of the anaerobic digestion microbiome can either take place via a non-targeted holistic evaluation of the microbial community through fingerprinting or by targeted monitoring of selected taxa. Here, we compared four different microbial community fingerprinting methods, i.e., amplicon sequencing, metaproteomics, metabolomics and cytomics, in their ability to characterise the full-scale anaerobic digestion microbiome. Cytometric fingerprinting through cytomics reflects a, for anaerobic digestion, novel, single cell-based approach of direct microbial community fingerprinting by flow cytometry. Three different digester types, i.e., sludge digesters, digesters treating agro-industrial waste and dry anaerobic digesters, each reflected different operational parameters. The α-diversity analysis yielded inconsistent results, especially for richness, across the different methods. In contrast, β-diversity analysis resulted in comparable profiles, even when translated into phyla or functions, with clear separation of the three digester types. In-depth analysis of each method's features i.e., operational taxonomic units, metaproteins, metabolites, and cytometric traits, yielded certain similar features, yet, also some clear differences between the different methods, which was related to the complexity of the anaerobic digestion process. In conclusion, cytometric fingerprinting through flow cytometry is a reliable, fast method for holistic monitoring of the anaerobic digestion microbiome, and the complementary identification of key features through other methods could give rise to a direct interpretation of anaerobic digestion process performance.

The integration of microbial technologies within the framework of the water-energy nexus has been taking place for over a century, but these mixed microbial communities are considered hard to deal with 'black boxes'. Process steering is mainly based on avoiding process failure by monitoring conventional parameters, e.g., pH and temperature, which often leads to operation far below the intrinsic potential. Mixed microbial communities do not reflect a randomised individual mix, but an interacting microbiological entity. Advance monitoring to obtain effective engineering of the microbiome is achievable, and even crucial to obtain the desired performance and products. This can be achieved via a top-down or bottom-up approach. The top-down strategy is reflected in the microbial resource management concept that considers the microbial community as a well-structured network. This network can be monitored by means of molecular techniques that will allow the development of accurate and quick decision tools. In contrast, the bottom-up approach makes use of synthetic cultures that can be composed starting from defined axenic cultures, based on the requirements of the process under consideration. The success of both approaches depends on real-time monitoring and control. Of particular importance is the necessity to identify and characterise the key players in the process. These key players not only relate with the establishment of functional conversions, but also with the interaction between partner bacteria. This emphasises the importance of molecular (screening) techniques to obtain structural and functional insights, minimise energy input, and maximise product output by means of integrated microbiome processes.

Temperature plays crucial roles in microbial interactions that affect the stability and performance of anaerobic digestion. In this study, the microbial interactions and their succession in the anaerobic digestion process were investigated at three levels, represented by (1) present and (2) active micro-organisms, and (3) gene expressions under a temperature gradient from 25 to 55 °C. Network topological features indicated a global variation in microbial interactions at different temperatures. The variations of microbial interactions in terms of network modularity and deterministic processes based on topological features, corresponded well with the variations of methane productions, but not with temperatures. A common successional pattern of microbial interactions was observed at different temperatures, which showed that both deterministic processes and network modularity increased over time during the digestion process. It was concluded that the increase in temperature-mediated network modularity and deterministic processes on shaping the microbial interactions improved the stability and efficiency of anaerobic digestion process.

Summary Several needs in the context of the water–energy–food nexus will become more prominent in the next decades. It is crucial to delineate these challenges and to find opportunities for innovative microbial technologies in the framework of sustainability and climate change. Here, we focus on four key issues, that is the imbalance in the nitrogen cycle, the diffuse emission of methane, the necessity for carbon capture and the deterioration of freshwater reserves. We suggest a set of microbial technologies to deal with each of these issues, such as (i) the production of microbial protein as food and feed, (ii) the control of methanogenic archaea and better use of methanotrophic consortia, (iii) the avoidance of nitrification and (iv) the upgrading of CO2 to microbial bioproducts. The central message is that instead of using crude methods to exploit microorganisms for degradations, the potentials of the microbiomes should be used to create processes and products that fit the demands of the cyclic market economy.

Anaerobic digestion is a well-established microbial-based technology for the treatment of organic waste streams and subsequent biogas recovery. A robust and versatile microbial community to ensure overall stability of the process is essential. Four full-scale anaerobic digestion plants were followed for one year to link operational characteristics with microbial community composition and structure. Similarities between digesters, community dynamics and co-occurrence between bacteria and archaea within each digester were analysed. Free ammonia concentration (>200 mg N L-1) and conductivity (>30 mS cm-1) hindered acetoclastic methanogenesis by Methanosaetaceae. Thus, methanogenesis was pushed to the hydrogenotrophic pathway carried out by Methanobacteriales and Methanomicrobiales. Firmicutes dominated the overall bacterial community in each of the digesters (>50%), however, principal coordinate analysis of Bray-Curtis indices showed that each of the four digesters hosted a unique microbial community. The uniqueness of this community was related to two phylotypes belonging to the Syntrophomonas genus (Phy32 and Phy34) and to one unclassified bacterium (Phy2), which could both be considered marker populations in the community. A clear differentiation in co-occurrence of methanogens with several bacteria was observed between the different digesters. Our results demonstrated that full-scale anaerobic digestion plants show constant dynamics and co-occurrence patterns in function of time, but are unique in terms of composition, related to the presence of marker populations.

Anaerobic digestion is shifting from a single-purpose technology for renewable energy recovery from organic waste streams to a process for integrated resource recovery. The valorisation of high-rate energy- and phosphorus-rich sludge creates the opportunity for their combined recovery. This phosphate is present in a precipitated form in the sludge, and its release into the liquid phase is an important issue before recovery can be achieved. The objective of this research was to exploit the “unwanted” sulphate reduction process for the release of phosphate into the liquid phase during anaerobic digestion, thus, making it available for recovery. Two different treatments were considered,i.e., a control digester and a digester to which sulphate was added, each operated in triplicate for a period of 119 days. The control digester showed stable methane production at 628 ± 103 mL CH4L−1d−1, with a feedstock COD (chemical oxygen demand) conversion efficiency of 89.5 ± 14.6 %. In contrast, the digester with sulphate addition showed a 29.9 ± 15.3 % decrease in methane production, reaching an “inhibited steady state”, but phosphate release into the liquid phase increased with a factor 4.5, compared to the control digester. This inhibited steady state coincided with a clear shift from a Methanosaetaceae to a Methanosarcinaceae dominated methanogenic community. Overall, the sulphate reduction process allows phosphate release during the anaerobic digestion process, yet, at the cost of a reduced methane production rate.

SCOPE: High red meat and sucrose consumption increases the epidemiological risk for chronic diseases. Mechanistic hypotheses include alterations in oxidative status, gut microbiome, fat deposition, and low-grade inflammation. METHODS AND RESULTS: For 2 weeks, 40 rats consumed a diet high in white or red meat (chicken-based or beef-based cooked mince, respectively), and containing corn starch or sucrose in a 2 × 2 factorial design. Lard was mixed with lean chicken or beef to obtain comparable dietary fatty acid profiles. Beef (vs chicken)-fed rats had higher lipid oxidation products (malondialdehyde, 4-hydroxy-2-nonenal, and hexanal) in stomach content and blood, and lower blood glutathione. Sucrose (vs corn starch)-fed rats showed increased blood lipid oxidation products and glutathione peroxidase activity, higher liver weight and malondialdehyde concentrations, and mesenterial and retroperitoneal fat accumulation. Beef-sucrose-fed rats had increased cardiac weight, suggesting pathophysiological effects on the cardiovascular system. The colonic microbiome of beef-sucrose-fed rats showed an outgrowth of the sulfate-reducing family of the Desulfovibrionaceae, and lower abundance of the Lactobacillus genus, indicating intestinal dysbiosis. Blood C-reactive protein, a marker for inflammation, was not different among groups. CONCLUSIONS: Consumption of a cooked beef-based meat product with sucrose increased oxidative stress parameters and promoted cardiac hypertrophy and intestinal dysbiosis. ispartof: Molecular Nutrition & Food Research vol:63 issue:2 ispartof: location:Germany status: published

Mechanisms explaining epidemiological associations between red (processed) meat consumption and chronic disease risk are not yet elucidated, but may involve oxidative reactions, microbial composition alterations, inflammation and/or the formation of toxic bacterial metabolites. First, in vitro gastrointestinal digestion of 23 cooked beef-lard minces, to which varying doses of nitrite salt (range 0–40 g/kg) and sodium ascorbate (range 0–2 g/kg) were added, showed that nitrite salt decreased protein carbonylation up to 3-fold, and inhibited lipid oxidation, demonstrated by up to 4-fold lower levels of ‘thiobarbituric acid reactive substances’, 32-fold lower 4-hydroxynonenal, and 21-fold lower hexanal values. The use of ascorbate increased the antioxidant effect of low nitrite salt levels, whereas it slightly increased protein carbonylation at higher doses of nitrite salt. The addition of a low dose of ascorbate without nitrite salt slightly promoted oxidation during digestion, whereas higher doses had varying antioxidant effects. Second, 40 rats were fed a diet of cooked chicken- or beef-lard minces, either or not cured, for three weeks. Beef, compared to chicken, consumption increased lipid oxidation (2- to 4-fold) during digestion, and gut protein fermentation (cecal iso-butyrate, (iso-)valerate, and fecal indole, cresol), but oxidative stress and inflammation were generally not affected. Cured, compared to fresh, meat consumption significantly increased stomach protein carbonylation (+16%), colonic Ruminococcaceae (2.1-fold) and cecal propionate (+18%), whereas it decreased cecal butyrate (-25%), fecal phenol (-69%) and dimethyl disulfide (-61%) levels. Fecal acetaldehyde and diacetyl levels were increased in beef-fed rats by 2.8-fold and 5.9-fold respectively, and fecal carbon disulfide was 4-fold higher in rats consuming cured beef vs. fresh chicken. Given their known toxicity, the role of acetaldehyde and carbon disulfide in the relation between meat consumption and health should be investigated in future studies.

Temperature is crucial for the performance of anaerobic digestion process. In this study of anaerobic digestion of swine manure, the relationship between the microbial gene expression and methane production at different temperatures (25–55 °C) was revealed through metatranscriptomic analysis. Daily methane production and total biogas production increased with temperature up to 50 °C, but decreased at 55 °C. The functional gene expression showed great variation at different temperatures. The function centralization (opposite to alpha-diversity), assessed by the least proportions of functional pathways contributing for at least 50% of total reads positively correlated to methane production. Temperature regulated methane production probably through reducing the diversity of functional pathways, but enhancing central functional pathways, so that most of cellular activities and resource were invested in methanogenesis and related pathways, enhancing the efficiency of conversion of substrates to methane. This research demonstrated the importance of function centralization for efficient system functioning.

Microbial management in anaerobic digestion is mainly focused on physically present and metabolically active species. Because of its complexity and operation near the thermodynamic equilibria, it is equally important to address functional regulation, based on spatial organisation and interspecies communication. Further establishment of the knowledge on microbial communication in anaerobic digestion through quorum sensing and nanowires is needed. Methods to detect centres of concentrated activity, related to the presence of highly active and well-connected species that take a central role in the anaerobic digestion process, have to be optimized. Bioaugmentation could serve as a crucial tool to introduce keystone species that may create or sustain such centres. Functional stability can be maintained by keeping the microbial community active. This results in a clear trade-off between functionally active and redundant microorganisms as primary basis for microbial community organization. Finally, a microbial community based prediction strategy for advanced process control is formulated.

Microbial protein (MP) can alleviate the increasing pressure of food demand on agriculture and our environment. For its sustainable production, feedstocks such as biomethane or (bio)hydrogen are needed. Here, we coupled biogas produced from agricultural waste directly with electrochemical biogas upgrading to subsequently produce MP from methane, hydrogen or a mixture thereof. Biogas was produced from co-digestion of pumpkin and pig manure at production rates of 0.73 ± 0.24 Lbiogas L−1reactor day−1 (59% CH4) and 0.59 ± 0.29 Lbiogas L−1reactor day−1 (50% CH4). The biogas was directed to the cathode of an electrochemical cell. At current densities of 20 and 40 A m−2, CO2 removal efficiencies of 88 ± 14% and 99 ± 1% were achieved. Enrichments of MP (hydrogen- and methane oxidizing bacterial cultures) were cultivated on either raw biogas or gases obtained from the cathode (CH4, CO2, H2) and anode (CO2, O2) in batch mode with external supplementation of O2 and H2 when required. The best performance was obtained when the cathode off-gas was used in terms of biomass concentration (0.585 g CDW L−1), yield (0.150 g CDW g−1 COD), efficiency of COD conversion to protein (17%) volumetric biomass productivity (0.226 g CDW L−1 day−1) and volumetric protein productivity (0.181 g protein L−1 day−1). The protein content was similar when using anode and cathode off-gases (66.3 ± 7.3% of CDW) with raw biogas resulting in a 6% lower protein content. This proof of concept demonstrated that electrochemical biogas upgrading enables steering MP production.